HepG2胰岛素抵抗模型的建立及在活性筛选中的应用

	HepG2胰岛素抵抗模型的建立及在活性筛选中的应用; Establishment of HepG2 insulin resistance model and its application in activity evaluation
	徐静悦
导师	胡江苗
摘要	Insulin resistance (IR) is not only the central of type 2 diabetes mellitus (T2DM), but also the soil that metabolic diseases such as hypertension, hyperlipidemia, and atherosclerosis. In recent years, it has received more and more attention. The research on IR mainly relies on the establishment of in vivo and in vitro model. From the perspective of molecular mechanisms, the establishment of stable and feasible in vitro cell models can not only shorten the modeling period but also reflect the situation of cellular insulin resistance more intuitively and clearly. In this subject, HepG2 cells were used as the object, and different insulin concentrations and different action time were used to induce cells to establish an insulin resistance model. This model was applied to evaluated the activity of the polysaccharide MSP-1 from the fruiting body of Morchella sextelata and 24 compounds from Dendrobium candidum. Screened out the compounds with the best activity of improvement of insulin resistance, and explored it’s mechanism at the molecular level. It is hoped to provide new ideas for finding more natural products that can improve insulin resistance. This paper is divided into five chapters, which are mainly discussed as follows: The first chapter reviews the background of insulin resistance, the activity of polysaccharides from Morchella and compounds from Dendrobium Provide a theoretical basis for subsequent research. In the second chapter is the extraction ,separation and structural of MSP-1. Using boilling water and ethanol precipitation, purification with DEAE-52 cellulose chromatography column, Sephadex S-200, Sephadex G-75 gelfiltration column, combined with HPLC-ELSD, infrared spectrum, PMP pre-column derivatization, etc techniques to detected the homogeneity, molecular weight, monosaccharide composition and structure of MSP-1. The results showed that MSP-1 was a homogeneous polysaccharide with a molecular weight of 1.17×104 Da. It was composed of 5 kinds of monosaccharides, such as mannose, rhamnose, galactose, xylose or arabinose, with a molar ratio of 23.3: 1: 1.5: 12.6. And has β-type glycosidic bond mannose residue and pyranose ring. The third chapter is the establishment of HepG2 insulin resistance model. By adjusting the concentration of insulin (10-4 mol/L-10-10 mol/L) and action time (12 h, 24 h, 36 h, 48 h, 60 h) to induce HepG2, use MTS method to detect cell viability and GOD-PAD method to detect glucose Consumption, the group with the highest cell viability and the least glucose consumption was selected as the best modeling condition. Using oil red O staining for morphological analysis and observation of lipid droplet distribution, detection of TG content, and analysis the stability and feasibility of the modeling conditions. The results showed that the best modelling conditions were 10-5mol/L for 24 hours. Glucose and lipid metabolism disorder appeared in the HepG2 at this condition, suggesting that the model was successful, and t
	2020-05
文献类型	学位论文
条目标识符	http://ir.kib.ac.cn/handle/151853/74197
专题	昆明植物所硕博研究生毕业学位论文
推荐引用方式 GB/T 7714	徐静悦. HepG2胰岛素抵抗模型的建立及在活性筛选中的应用, Establishment of HepG2 insulin resistance model and its application in activity evaluation[D],2020.

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